MS based peptide and protein sequencing via reactions of lysine residues with peroxycarbonate compounds
a technology of lysine residues and peroxycarbonate compounds, applied in the field of proteomics and analytical methods, can solve the problems of time-consuming and difficult separation and identification process of proteins that include enzymatic digestion, and achieve the effect of convenient and rapid method of quantifying proteins and high through-the-procedur
Inactive Publication Date: 2010-12-28
VANDERBILT UNIV
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Benefits of technology
The present invention provides a method for predictably fragmenting large proteins in a mass spectrometer and identifying protein sequences using peroxycarbonate compounds. This method is efficient and reliable, and can be performed using different types of mass spectrometers. The invention also provides novel peroxycarbonate / peptide and peroxycarbonate / protein combinations for chemical modification of peptides and proteins. Overall, the invention offers a convenient and rapid way to quantify proteins and peptides.
Problems solved by technology
As stated above, the prior art process of separating and identifying proteins that includes enzymatic digestion is a difficult and time-intensive process.
Method used
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[0021]Digestion by protease enzymes is an important part of the standard protocol used to provide protein primary structural information. Large proteins are converted in a predictable way to smaller peptide fragments that can be readily identified. Thus, the cornerstone methods used in protein identification are enzymatic proteolysis, separations such as chromatography and electrophoresis, and analysis of isolated small peptide fragments by mass spectrometry.
[0022]Chemical digestion of proteins has also provided strategies that are helpful in assignment of peptide sequences. Both chemical and enzymatic protein digestion, however, can be cumbersome and time-consuming and attempts to avoid this step by the use of powerful mass spectrometric “top-down” approaches are being explored to provide protein primary sequences.
[0023]Strategies that combine in one step a predictable chemical-based protein digestion with mass spectrometry might prove to be convenient for rapid primary peptide seq...
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Abstract
A method of modifying protein samples that comprises combining the sample with a peroxycarbonate solution and inserting the sample into a mass spectrometer. The present invention also includes methods of N-terminus characterization.
Description
TECHNICAL FIELD OF THE INVENTION[0001]The present invention relates to the field of proteomics and analytical methods used in the determination of protein structures. More particularly, the present invention provides a process for modifying peptides and proteins on lysine amino groups such that the modification leads to fragmentation at or near the lysine position in a mass spectrometer.[0002]Additionally, the present invention relates to methods of modifying amino acids and peptides. More particularly, embodiments of this method modify amino acids and peptides to form a t-butyl-peroxycarbamate compound. This aspect of the present invention is useful for the development of N-terminus characterization methods.BACKGROUND OF THE INVENTION[0003]A popular method for sequencing proteins (such as, for example, in proteomics applications) involves enzymatic digestion (trypsin, chymotrypsin, etc.) of a large protein into smaller peptides and subsequent MS sequencing of the smaller peptides.[...
Claims
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IPC IPC(8): G01N24/00G01N33/00C07K
CPCC07C409/38C07C409/40G01N33/6851C07K1/128Y10T436/24
Inventor PORTER, NED A.MASTERSON, DOUGLAS S.YIN, HUIYONGCAPRIOLI, RICHARD M.NORRIS, JEREMY L.
Owner VANDERBILT UNIV